Abstract
The muscle isozyme of glycogen phosphorylase (MGP)is developmentally and neurally regulated in rat muscle. A series ot deletion derivatives were generated in a CAT reporter vector to identify elements in the 5' noncoding region of MGP required for developmental regulation. A 269 bp fragment was identified that conferred 15-fold higher levels of CAT activity in C2C12 myotubes than myoblasts. This fragment contained four E box motifs located at -167, -118, -18 and +29, a CArG-like element (CCAAAATAGC) at -109, a GC rich region at -63 and a TATA motif 27 bp 5' of the transcriptional start point. Point mutations of the E boxes had little affect on the developmental regulation of MGP. Three of the muscle regulatory factor proteins (MyoD, myogenin and MRF4) transactivated the MGP promoter by only 2-fold and one them ( myf-5) had no effect on CAT activities in cotransfection studies in the nonmuscle cell line C3H10T1/2. Deletion of the CArG-like sequence reduced the difference between MT and MB CAT activity to only 1.8-fold. This may be a potential area of importance in the developmental-specific regulation of MGP in muscle cells. Further deletion of the GC motif eliminated any measurable CAT activity and defined the minimal promoter region between -76 and +3.
| Original language | English (US) |
|---|---|
| Journal | FASEB Journal |
| Volume | 10 |
| Issue number | 3 |
| State | Published - 1996 |
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ASJC Scopus subject areas
- Agricultural and Biological Sciences (miscellaneous)
- Biochemistry, Genetics and Molecular Biology(all)
- Biochemistry
- Cell Biology
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Developmental expression of the rat muscle glycogen phosphorylase gene is not directly regulated by the four muscle regulatory factor proteins. / Froman, B.; Gorin, Fredric A.
In: FASEB Journal, Vol. 10, No. 3, 1996.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Developmental expression of the rat muscle glycogen phosphorylase gene is not directly regulated by the four muscle regulatory factor proteins
AU - Froman, B.
AU - Gorin, Fredric A
PY - 1996
Y1 - 1996
N2 - The muscle isozyme of glycogen phosphorylase (MGP)is developmentally and neurally regulated in rat muscle. A series ot deletion derivatives were generated in a CAT reporter vector to identify elements in the 5' noncoding region of MGP required for developmental regulation. A 269 bp fragment was identified that conferred 15-fold higher levels of CAT activity in C2C12 myotubes than myoblasts. This fragment contained four E box motifs located at -167, -118, -18 and +29, a CArG-like element (CCAAAATAGC) at -109, a GC rich region at -63 and a TATA motif 27 bp 5' of the transcriptional start point. Point mutations of the E boxes had little affect on the developmental regulation of MGP. Three of the muscle regulatory factor proteins (MyoD, myogenin and MRF4) transactivated the MGP promoter by only 2-fold and one them ( myf-5) had no effect on CAT activities in cotransfection studies in the nonmuscle cell line C3H10T1/2. Deletion of the CArG-like sequence reduced the difference between MT and MB CAT activity to only 1.8-fold. This may be a potential area of importance in the developmental-specific regulation of MGP in muscle cells. Further deletion of the GC motif eliminated any measurable CAT activity and defined the minimal promoter region between -76 and +3.
AB - The muscle isozyme of glycogen phosphorylase (MGP)is developmentally and neurally regulated in rat muscle. A series ot deletion derivatives were generated in a CAT reporter vector to identify elements in the 5' noncoding region of MGP required for developmental regulation. A 269 bp fragment was identified that conferred 15-fold higher levels of CAT activity in C2C12 myotubes than myoblasts. This fragment contained four E box motifs located at -167, -118, -18 and +29, a CArG-like element (CCAAAATAGC) at -109, a GC rich region at -63 and a TATA motif 27 bp 5' of the transcriptional start point. Point mutations of the E boxes had little affect on the developmental regulation of MGP. Three of the muscle regulatory factor proteins (MyoD, myogenin and MRF4) transactivated the MGP promoter by only 2-fold and one them ( myf-5) had no effect on CAT activities in cotransfection studies in the nonmuscle cell line C3H10T1/2. Deletion of the CArG-like sequence reduced the difference between MT and MB CAT activity to only 1.8-fold. This may be a potential area of importance in the developmental-specific regulation of MGP in muscle cells. Further deletion of the GC motif eliminated any measurable CAT activity and defined the minimal promoter region between -76 and +3.
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UR - http://www.scopus.com/inward/citedby.url?scp=33749298955&partnerID=8YFLogxK
M3 - Article
AN - SCOPUS:33749298955
VL - 10
JO - FASEB Journal
JF - FASEB Journal
SN - 0892-6638
IS - 3
ER -